Reciprocal effects in anther cultures of wheat hybrids

This study was conducted to determine the reciprocal effects for anther culture response in wheat (Triticum aestivum L.) using a set of 4 × 4 full diallel crosses. Both reciprocal and nuclear genetic effects were highly significant for anther culture response and useful for selection and breeding purposes. General combining ability (GCA) effects were predominant for all investigated anther culture traits. Also, significant differences for specific combining ability (SCA) effects were detected between reciprocal crosses. Although significant reciprocal differences for responding anther, callus number and green plant regeneration were recorded in some reciprocal crosses, there were no significant reciprocal differences for albino plant regeneration. The use of one parent as male or female could lead to change at the production of green plants from the F₁ hybrids and screening of inbred lines for response to anther culture, without reciprocal effects, could decrease the utilization of breeding material.     

Genetic studies on tissue culture response and some agronomical traits in Egyptian bread wheat

The main effort of wheat breeder is the detection of genes and to merge them in a particular genotype using most suitable combination. Five Egyptian cultivars of bread wheat (Triticum aestivum L.) were crossed in a half diallel mating design to produce 10 crosses. The genetic potential of embryogenic callus (EC%), plant regeneration (RGP%) response and its association with heading date (HD) and grain yield per plant (GY/P) were investigated. The results showed that GY/P was significantly and positively correlated with EC% and RGP%. The combining ability analysis showed that the magnitudes of general combining ability (GCA) were higher than those of specific combining ability (SCA) for both tissue culture response and agronomic traits. The promising crosses which exhibited desirable SCA effects, showed also high useful heterosis for all studied traits. The magnitudes of additive genetic variance (σ²A) were larger than those of non-additive ones (σ²D) for all studied traits except for number of days to heading. The estimates of narrow sense heritability were 84.56%, 82.13%, 43.46% and 70.28% for the percentage of EC%, RGP%, HD and GY/P, respectively. The genetic similarity percents based on RAPD markers ranged from 76% to 93% between the cultivars. The UPGMA cluster analysis revealed that the cultivars could be divided into two main clusters. The range of Euclidean distances based on morphological characters among the cultivars was relatively wide (4.37–27.87), indicating relatively high amount of phenotypic variation. A significant positive correlation between Euclidean distance and RAPD distance (0.72⁷) was found.     

Inheritance of somatic embryogenesis and organ regeneration from immature embryo cultures of winter wheat

Summary Diallel analyses of F₁ and reciprocal crosses among five winter wheat lines show that additive, non-additive, and cytoplasmic genetic effects were significant in the genetic control of somatic embryogenesis, shoot, and root induction frequencies as well as in numbers of somatic embryos, shoots, and roots. However, additive genetic effect appears to be most important since, in most cases a larger portion of the cross variation was accounted for by general combining ability. The results suggest that somatic embryogenesis and organ regeneration in winter wheat can be improved through genetic manipulation. Due to the presence of maternal effects, it may be critical to use a suitable genotype as a female parent in a selection program.Contribution of the College of Agricultural Sciences, Texas Tech University, Journal No. T-4-266     

Inheritance of anther culture derived green plantlet regeneration in wheat (Triticum aestivum L.)

A study was set up to determine the inheritance and combining ability of the factors anther culture response and green plant regeneration. Reciprocal crosses were made between cultivar Ringo Sztar, showing high anther culture response and the cultivars Ciano 067 and Benoist H77022, showing a high level of green plant regeneration. Averaged over all genotypes, 23.0% of the anthers responded and a callus induction frequency of 77.8% was observed. Of all the embryos, 43.0% developed into plantlets, 25.6% of the regenerants being green, the result being that 3.3 green plants per 100 anthers were formed. Genotypic effects accounted for 57.7%, 86.3% and 77.5% of the total variance of anther culture response, callus induction frequency and embryo induction frequency, respectively. Additive and dominant gene action was detected for all characteristics, including green plant regeneration. No reciprocal differences were found for anther culture response, embryo induction frequency and green plant regeneration, indicating no cytoplasmic effects. A small but significant reciprocal difference was found for callus induction frequency. Embryo production was primarily correlated with anther culture response and not with the number of embryos produced per plated anther or per responding anther. Possible mechanisms for the inheritance of green plant regeneration are discussed.Abbreviations CIRA callus induction frequency per responding anther - ERA embryo induction frequency per responding anther - FHB fusarium head blight - MS-medium Murashige & Skoog (1962) medium - REML residual maximum likelihood     

Selection of stable mutants from cultured rice anthers treated with ethyl methane sulfonic acid

To increase the frequency of stable mutants from cultured anthers of rice, the effects of EMS treatment on callus induction, plant regeneration and mutant induction were investigated according to the timing of treatment after anther inoculation on the medium. The frequency of callus induction was highest in anthers treated with 0.5% EMS 10 days after culture. Anthers treated directly at the initiation of culture exhibited a very low callus induction level, and the such calluses exhibited a poor plant regeneration capacity. The frequency of regeneration of green plants was significantly decreased by EMS treatments immediately after anther inoculation as compared with control. The frequencies of stable mutants were 20.7% and 12.0% in EMS treatments at 10 and 20 days, but unstable mutants were 43.1% and 52.6%, respectively. A total of 14 stable mutants, semidwarf mutants (4 lines), grain-shape mutants (2 lines) and glabrous mutants (8 lines) were selected from doubled haploid lines of the A₂ generation. The frequencies of callus induction, green plant regeneration and stable mutants were maximal in anthers treated with 0.5% EMS 10 days after culture.     

Genetic studies of rice (Oryza sativa L.) anther culture response

Anthers of two rice (Oryza sativa L.) varieties, Lunhui 422 (P₁) and Jinzao 5 (P₂), their F₁, F₂ and first backcross generation-F₁ x Lunhui 422 (B₁), and F₁ x Jinzao 5 (B₂)-were cultured in L8 medium to study the inheritance of rice anther culturability using generation mean analysis. Significant effects of generation were observed for the four traits measured: anther response (%), frequency of callus induction (%), frequency of callus differentiation (%) and culture efficiency (%). Variation among the generations was similar for all traits: significant differences were found among six generations and the means of the P₂ and B₂ were significantly lower than those of the other generations. The frequency of callus differentiation showed a nonsignificant difference among the P₁, F₁, F₂ and B₁ generations which had slightly high values than the P₂ and B₂. Additive genetic variance (V_A) was higher than non-additive genetic variance (V_D) for anther response and frequency of callus induction. However, A_V was lower than V_D for frequence of callus differentiation and culture efficiency, V_A was significant for all traits except for the culture efficiency, and V_D was nonsignificant for all traits except for the frequency of callus differentiation. On the other hand, environmental variation (V_E) was significant for the 4 traits. Narrow-sense heritability estimates were 95.52%, 82.19% and 13.54% for anther response, frequency of callus induction and culture efficiency, respectively.Abbreviations 2,4-d 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid     

Genetic studies of anther culture ability in rice (Oryza sativa)

Inheritance of three anther and culture characters, callus induction, green plant regeneration and culture efficiency was studied using incomplete diallel crosses with a gamete model. It was suggested that callus induction was mainly controlled by gametic additive effects and with less effect of the maternal effects. Green plant regeneration was mainly determined by maternal effects with less influence of gametic additive effects. Culture efficiency was controlled by gametic additive, maternal and cytoplasmic effects. Cultivar Lunhui 422 showed positive genetic effects for all three traits and was a very good parent for rice anther culture breeding. Significant positive heterosis was observed for callus induction. Both gametic additive and maternal correlations contributed to the significant genotypic and phenotypic correlations between callus induction and green plant regeneration suggesting these two traits to be linked.Abbreviations 2,4-d 2,4-dichlorphenoxyacetic acid - NAA -napthaleneacetic acid     

Effects of light regimes on anther culture response in bread wheat

This experiment was initiated to further test the effects of light regimes during callus induction and plant regeneration on anther culture response of spring wheat (Triticum aestivum L.). Spring wheat cultivars 'Edwall' and 'WA 7176' with high callus induction from anther culture but low green plant production were used. Different gro-lux light and dark regimes during callus induction, and gro-lux light and fluorescent light regimes during plant regeneration were used. Callus induction decreased significantly at relatively high light intensity (315 μmol m⁻² s⁻¹) applied at any period of culture when compared to continuous dark. Light regimes used continuously and from the 15^th to the last day of callus induction also had a significant negative effect on plant regeneration compared to continuous dark and light application in the first half of callus induction. During plant regeneration, '15 day dark + 7 day gro-lux light' significantly increased plant regeneration compared to both 'gro-lux' and 'fluorescent light' regimes. Light regimes during both callus induction and plant regeneration and their interaction effects were found to be highly significant on green plant proportion and green plant yield. 'Continuous light' application during callus induction increased green plant proportion more than other applications in contrast to its negative effect on plant regeneration. During plant regeneration, '15 day dark + 7 day gro-lux light' had the higher green plant proportion compared to only 'fluorescent light' and only 'gro-lux light'. The highest green plant yields were obtained from '15 day dark + 7 day gro-lux light' during plant regeneration in combination with either 'continuous dark' or 'continuous light' regimes during callus induction. This revised version was published online in June 2006 with corrections to the Cover Date.     

Screening wheat genotypes for high callus induction and regeneration capability from anther and immature embryo cultures

Plant regeneration via tissue culture varies with the genotype and is an important factor in establishing cell selection and genetic transformation systems. To select genotypes – especially Japanese ones – with a high regeneration capability, we screened 107 wheat genotypes (78 domestic, 29 foreign) for callus induction and regeneration capability from anther and immature embryo cultures. For anther culture, 83 of 107 genotypes tested induced calli and 45 regenerated plants. Only 9 genotypes, however, produced green plants, 25 produced only albino plants, and 11 produced both green and albino plants. Glennson 81 was the highest in callus induction, followed by Orofen, Danchi–komugi and Chris. The genotypes with a relatively high regeneration capability were Framala 80 at 24% and Glennson 81 at 19%, these two genotypes produced only green plants. For immature embryo culture, 97 genotypes showed a 90% callus induction rate and 74 genotypes regenerated plants. Very few genotypes produced albino plants. The genotypes with a high regeneration capability were Genaro 81 at 90%, Chinese Spring at 80%, and Norin 75 at 75%. This revised version was published online in June 2006 with corrections to the Cover Date.     

杂交水稻主要农艺性状配合力遗传力分析

选用生产上常用的5个杂交水稻不育胞质与5个高配合力恢复系,采用NCII模式对产量性状、芒、粒形等主要农艺性状的亲本配合力、方差贡献率及遗传力进行分析。结果表明：胞质效应和恢复系效应在所测性状中均达到显著或极显著差异水平,芒长、粒形、单穗重受环境的影响较小;产量和收获指数的非加性遗传作用明显,受环境效应的影响也较大。亲本效应方面,恢复系的效应远大于胞质效应;产量性状JW型（爪哇型）胞质和蜀恢527的一般配合力效应较高,同时JW型胞质和R21属于Ⅰ类亲本;粒形性状G型（冈型）胞质和蜀恢527的一般配合力效应最高,同时均属于Ⅰ类亲本。     

Response of flax genotypes to doubled haploid production

Forty-four flax genotypes with a diverse genetic background were evaluated for anther culture response using a standard anther culture protocol in order to determine the feasibility to initiate a routine haploid production system in applied breeding programs. A strong genotype effect on callus induction and shoot regeneration in anther culture was found in this study. A number of genotypes, including two low cadmium content lines 96-11785 and 96-11826, a high oil content line 96-22109 and a high linolenic acid content line M 4919 were identified as highly responsive. The impact of the findings in this study on flax breeding was discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

日照长短和脯氨酸处理对光敏核不育水稻花药培养的影响

本试验对粳型光敏核不育系Ｎ５０４７Ｓ、７００１Ｓ、Ｎ５０８８Ｓ和ＨＰ１３Ｓ、常规粳稻品种渐农大４０和０２４２８及由它们所配制的Ｆ１杂种进行花药培养,主要结果如下：不同光敏核不育系及其杂种Ｆ１出愈率、绿苗率和培养力存在明显差异;光敏核不育系的花培效率并不一定比非光敏核不育系低;无论光敏不育系还是非光敏核不育系,长日都比短日处理具更高的愈伤组织诱导率,光敏核不育系长日下生长取样接种,脱分化培养基中添加     

Joint modeling of additive and non-additive genetic line effects in single field trials

A statistical approach is presented for selection of best performing lines for commercial release and best parents for future breeding programs from standard agronomic trials. The method involves the partitioning of the genetic effect of a line into additive and non-additive effects using pedigree based inter-line relationships, in a similar manner to that used in animal breeding. A difference is the ability to estimate non-additive effects. Line performance can be assessed by an overall genetic line effect with greater accuracy than when ignoring pedigree information and the additive effects are predicted breeding values. A generalized definition of heritability is developed to account for the complex models presented.     

芍药花药愈伤组织诱导及体细胞胚发生

以芍药(Paeonia lactiflora)品种粉玉奴花药为外植体,研究不同浓度2,4-D对愈伤组织诱导、体胚发生及植株再生的影响,采用组织细胞学方法观察愈伤组织以及体细胞胚发育过程,采用根尖染色体法鉴定再生植株倍性。结果表明,芍药花药愈伤组织诱导的最适培养基为MS+1 mg·L–1 2,4-D+1 mg·L–1 N...     

粳稻花药培养基因型差异的研究

对7个粳稻(Oryza sativa L. sub sp. kcng)品种按双列杂交方式配成的21个杂交组合进行花药培养,测定它们的愈伤组织诱导率和绿苗分化率。统计分析表明,基因型变异方差、一般配合力方差及特殊配合力方差均达到极显著水平。对亲本的一般配合力效应和组合的特殊配合力效应也进行了估算。研究表明,粳稻花药培养力是可遗传的,用高培养力的材料做亲本以提高杂种花粉植株的产量是有效的。此外还观察到供体植株在小孢子单核晚期前10天左右遇到特殊低温会使愈伤组织诱导率下降。     

Effect of the 1B/1R translocation on anther culture ability in wheat (Triticum aestivum L.)

Using two varieties, their reciprocal hybrids, F₈ lines and doubled haploids, results confirmed that three genetic components are involved in wheat anther culture ability, viz embryo induction frequency, regeneration ability and the frequency of albinism. In these experiments, no significant maternal effects were noticed. For embryo yields, transgressive lines were obtained from hybrids between distant genotypes. Regeneration of green plants depended upon two independent traits: regeneration ability and the frequency of albinos. F₈ lines and two doubled haploids equaled the 50% regeneration rate of the hybrids, but they only regenerated green plants. Based upon cytological examination and gliadin patterns, it is suggested that genes favoring regeneration ability could be linked to the 1BL-1RS translocated chromosome from Aurora.Abbreviations DH doubled haploids - MS Murashige and Skoog - MPG multicellular pollen grains     

云南野生稻不同染色体组型和外植体材料的离体培养研究

云南野生稻不同外植体愈伤组织诱导能力差别较大。花粉培养中愈伤组织诱导率差异在0％～11．8％之间,用成熟胚诱导愈伤组织,其诱导率在18．0％～35．2％之间,茎叶培养则在12．0％～25．0％之间。云南野生稻不同外植体诱导的愈伤组织再分化为绿苗的分化率在8．3％～100．0％之间。疣粒稻组培特性最好,东乡普通野生稻和景洪普通野生稻次之,药用稻最难组培。本文建立了疣粒、东乡、景洪普野3种野生稻的离体无性系,为长期保存云南野生稻资源奠定了基础。     

Elevated carbon supply altered hypericin and hyperforin contents of St. John's wort (Hypericum perforatum L. cv `New Stem') grown in bioreactors

For the high frequency selection of salt-tolerant doubled haploids (DHs) using rice anther culture, the efficiency of anther culture was investigated with different genotype, media condition and NaCl concentrations. The six F₁ hybrids obtained by backcross or three-way cross between indica and japonica differed in salt tolerance. The efficiencies of callus induction and plant regeneration was decreased by NaCl concentration and salt tolerance of donor variety, and those in japonicas were higher than those in indicas. The percentages of callus induction in Gyehwa 5 (japonica, tolerant) and IR61633-B-2-2-1 (japonica, sensitive) were 21.1 and 13.5% on agar medium containing 0.3% NaCl, respectively. The plant regeneration of callus derived from anther floating culture in liquid media was less than that on solid medium. In four F₁ hybrids, the frequencies of high salt-tolerant DHs were 21.4 and 8.9% in 0.3% NaCl medium and the control, respectively. The high frequency of salt-tolerant DHs could be selected in the callus induction medium (0.3% NaCl) and in the combinations crossed with salt-tolerant japonica as the third parent.     

Induced androgenesis in tomato (Lycopersicon esculentum Mill). II. Factors affecting induction of androgenesis

The influence on androgenesis of donor plant growth conditions, anther size and developmental stage of the microspore, medium composition and different anther treatments prior to culture was investigated in L. esculentum Mill. cv Roma and its hybrids. Growth conditions of donor plants affected the induction of tomato androgenesis. Anthers isolated from plants grown in the greenhouse during winter at high humidity and in short days possessed higher androgenetic ability than those grown in the field. The physiological state and age of the donor plants also influenced the processes investigated. Regarding the developmental stage of microspores, the period from prophase to telophase II is optimal for tomato anther implantation. More then 20 culture media were tested. Two, based on Murashige and Skoog medium were selected as most favourable for callus induction, organogenesis and regeneration. The effect on callus induction of 2ip in combination with indole-3-acetic acid (IAA) was greater than that of zeatin and IAA. Zeatin promoted entire plant regeneration. A highly significant interaction between genotype and medium was observed. Temperature and gamma ray treatments of anthers enhanced callus production, shoot formation and plant regeneration. Treatments at 4 °C (48 h) and 10 °C (9 days) stimulated these processes. Combined treatment of anthers with 4 Gy and 10 °C for 9 days was the most efficient. Received: 5 September 1997 / Revision recieved: 5 June 1998 / Accepted: 15 June 1998     

Antimitotic and hormone effects on green double haploid plant production through anther culture of Mediterranean japonica rice

Rice double haploid (DH) plants are produced mainly through anther culture. In order to improve the anther culture protocol, microspores of two japonica rice genotypes (NRVC980385 and H28) were subjected to three growth regulator combinations and four colchicine treatments on induction medium. In addition, a post anther culture procedure using colchicine or oryzalin was tested to induce double haploid plantlets from haploid plantlets. A cold pre-treatment of microspores for 9 days at 10 °C increased callus induction 50-fold in the NRCV980385 genotype. For both genotypes, 2 mg L^?1 2,4-D and 1 mg L^?1 kinetin on colchicine-free induction medium gave the best culture responses. The culturability of both genotypes changed on colchicine-supplemented induction media. A high genotype dependency was recorded for callus induction, callus regenerating green plantlets and regeneration of green double haploid plantlets. Colchicine at 300 mg L^?1 for 48 h enhanced callus induction 100-fold in H28. Colchicine-supplemented media clearly improved green double haploid plantlet regeneration. We showed that the post-anther culture treatment of haploid plantlets at 500 mg L^?1 of colchicine permitted fertile double haploid plantlets to be generated. Finally, an enhanced medium-throughput flow cytometry protocol for rice was tested to analyse all the plantlets from anther and post anther culture.